Fig. 3: Developing L3 wing discs become increasingly hypoxic in environmental normoxia.
a-a’ Fluorescence from SyHREns in L3 discs increases during development in environmental normoxia. Representative images are shown in (a) (all discs imaged under identical conditions), and quantification of wing pouch fluorescence intensity is shown in (a’). Scale bar = 100 μm. n ≥ 7 for each time point, except for data at 96 h AEL where n = 6. After confirming normality of the data, statistical significance was assessed by a two-sided unpaired t test. b-b’ Fluorescence from HypoxyprobeTM adducts in L3 discs increases during development in environmental normoxia. 72 and 92 h AEL larvae were simultaneously reared on fly food containing 2 mg/mL HypoxyprobeTM for 24 h prior to fixation. Representative images are shown in (b) (all discs imaged under identical conditions), and quantification of wing pouch fluorescence intensity is shown in (b’). Scale bar = 100 μm. n = 12 for each time point. After confirming normality of the data, statistical significance was assessed by a two-sided unpaired t test. c-c’ Fluorescence from anti-V5 staining in L3 discs expressing FlHypox increases during development in normoxia. Representative images are shown in (c) (all discs imaged under identical conditions) and quantification of wing pouch fluorescence intensity in (c’). Scale bar = 100 μm. n ≥ 5 for each time point. After confirming normality of the data, statistical significance was assessed by a two-sided unpaired t test. d-d’ Fluorescence from anti-V5 staining in L3 discs expressing FlHypoxP850A decreases during development under environmental normoxia. Representative images are shown in (d) (all discs imaged under identical conditions) and quantification of wing pouch fluorescence intensity is shown in (d’). Scale bar = 100 μm. n ≥ 10 for each time point. After confirming normality of the data, statistical significance was assessed by a two-sided unpaired t test.
